Human gliomas aggressively invade the adjacent normal brain tissue to render surgical treatments for this disease impossible. Two different mouse models of glioma demonstrate two distinct patterns of migration and invasion of adjacent tissues: PDGF-induced oligodendrogliomas invade the brain as individual cells, while Ras-driven gliomas invade by tracking along blood vessels. This translational, bioengineering proposal will investigate these observations by developing and implementing a novel, nano-microfluidic assay to examine the mechanisms of brain tumor cell migration via real-time visualization of cell movement, and detection of spatial and temporal activity of signaling protein components. [unreadable] [unreadable] Our project will develop custom nanoprobes to bind, both, the extracellular and intracellular domains of the PDGF receptor in order to visualize its spatial distribution and temporal location during real-time migration in motile glioma. Further, our experiments will determine the subcellular localization of PDGFR domains relative to the direction of migration by using our microfluidic migration assay to impose controllable, one-dimensional ligand gradients onto individual cells. [unreadable] [unreadable] The proposed methodology is unique and high risk because it is the first to develop analytical tools able to measure translocations of intracellular proteins induced by a controlled, extracellular environment. Successful development of our proposed approach will enable future research to investigate cell migration via intracellular visualization of key proteins from receptors to Ras. Such ground-breaking results will elucidate unprecedented details of intracellular protein behavior during tumor cell dispersal. [unreadable] [unreadable]